Edelweiss Applied Science and Technology ISSN: 2576-8484 Vol. 9, No. 2, 853-862 2025 Publisher: Learning Gate DOI: 10.55214/25768484.v9i2.4619 © 2024 by the authors; licensee Learning Gate © 2025 by the authors; licensee Learning Gate History: Received: 7 December 2024; Revised: 24 January 2025; Accepted: 31 January 2025; Published: 6 February 2025 * Correspondence: j.borjac@bau.edu.lb Phytochemicals analysis of Arum palaestinum extracts and their antibacterial effect against Pseudomonas syringae pv. tomato (Pst) Samaher Ghaith1, Dany Abi Shahine2,3,4, Antoine Abou Fayad2,3, Michel Afram5, Jamilah Borjac1* 1Beirut Arab University, Department of Biological Sciences, Faculty of Science, Debbieh, Lebanon; j.borjac@bau.edu.lb (J.B.). 2Department of Experimental Pathology, Immunology and Microbiology, Faculty of Medicine, American University of Beirut, Lebanon. 3Center for Drug Discovery, American University of Beirut, Lebanon. 4Laboratory of Biodiversity and Functional Genomics, UR EGP, Faculty of Science, Université Saint-Joseph de Beyrouth, Beirut, Lebanon. 5General Director of Lebanese Agriculture Research Institute (LARI), Tel Amara, Lebanon. Abstract: Arum palaestinum is a traditional medicinal wild plant indigenous to the Levant. The present study explores its phytochemical constituents and antibacterial activity in agriculture to control and inhibit the growth of Pseudomonas syringae pv. tomato (Pst). Aqueous, ethanolic, and chloroform extracts of the dried leaves, flowers, and rhizomes were prepared at a final concentration of 50 mg/mL. HPLC-MS analysis of A. palaestinum extracts revealed the presence of 13 previously characterized compounds and 2 new ones. Using the disc diffusion method and compared to ciprofloxacin and copper oxychloride that were used as controls, the aqueous extracts of the leaves, flowers, and rhizomes exhibited significant inhibition against P. syringae with high inhibition zones (IZDs). Based on the broth dilution method, the aqueous extracts possessed the greatest antimicrobial effect against P. syringae, followed by the ethanolic extracts, which showed a moderate antimicrobial effect for each part. In the field, the aqueous and ethanolic extracts controlled and eradicated the speck disease on tomato better than the chloroform extracts. Therefore, A. palaestinum can be used as an alternative to control bacterial speck disease and deserves to be further studied in organic agriculture. Keywords: Antibacterial activity, Arum palaestinum, LC-MS, Organic agriculture, Pseudomonas syringae, Speck disease. 1. Introduction Araceae is a large plant family represented by 3800 species in 118 genera spreading over a wide range of ecological habitats starting from sea level to a 3000 m altitude [1]. Arum palaestinum Boiss. (Black Calla Lily or Louf, A. palaestinum ), is one of ~26 species of the Arum genus belonging to this family [2, 3]. It is native to Europe, Northern Africa, Western Asia, with the highest species diversity found in the Mediterranean region [4-6]. It grows up to 0.82 ft and blooms in the spring, between March and April. The plant is recognized by its dark purplish-black spadix enclosed in a reddish-brown spathe [7]. Black lily is a typical “cryptic” species, since its appendix emits mainly ethyl acetate, producing a smell of rotten fruit. In many countries, the aerial parts of A. palaestinum are considered ornamental plants, animal fodder and can be edible dried or after soaking in salty water. The plant is used in folk medicine to cure several chronic diseases such as stomach acidity, atherosclerosis, cancer, and diabetes [6, 8, 9]. Numerous studies have shown that A. palaestinum leaves contains several biologically active phytochemicals with the highest quantity of saponin, alkaloid, phenols and flavonoids [4, 6, 10]. Bacterial plants’ diseases are often very difficult to eradicate. Few effective strategies are adopted to control them. Medicinal plants are among the richest bioresources of the metabolites that are currently mailto:j.borjac@bau.edu.lb 854 Edelweiss Applied Science and Technology ISSN: 2576-8484 Vol. 9, No. 2: 853-862, 2025 DOI: 10.55214/25768484.v9i2.4619 © 2025 by the authors; licensee Learning Gate used to treat bacterial and other infections [11, 12]. Yield losses in food crops due to plant pathogenic bacteria are significant and increasing over the years. The increasing losses caused by bacterial plant pathology are explained by the emerging resistance of bacteria to the chemical agents used in plant protection. Moreover, these chemical agents harm the environment through residue accumulation leading to soil pollution and the perturbation of the soil's inner ecosystem [13]. The plant pathogenic Pseudomonas syringae (P. syringae) species is one of the most important plant pathogenic bacteria. It is divided into different pathovars based on their host range. A certain pathovar usually has a limited host range; however, the whole P. syringae group can cause diseases for a wide range of host plants. P. syringae pv. tomato (Pst) causes bacterial speck on tomato plants and is considered a highly aggressive pathogen once inside the plant [14, 15]. Among the many symptoms it causes, one can find brown- black leaf spots that are surrounded by chlorotic margin; dark superficial specks on green fruit where they can become sunken on ripe fruits, surrounded by a zone of delayed ripening. Stunting and yield loss, particularly if young plants are infected, will result due to the infection, thus reducing the market value of the speckled fruit [16, 17]. Many research tackled the chemical composition of A. palaestinum [4, 18-20]; however, the need for an extensive identification of its phytochemical components seems imperative. Undoubtedly, the agricultural use of this plant as a bio-bactericide has robustly prompted us to carry out the phytochemical analysis of this Lebanese wild plant as a promising antibacterial plant. For that reason, this study presents a comprehensive qualitative characterization of the phytochemical released in aqueous, ethanolic and chloroform extracts of A. palaestinum leaves, flowers and rhizomes using liquid chromatography–tandem mass spectrometry (LC-MS) and their activities against the pseudomonas syringae pv. tomato (Pst). 2. Materials and Methods 2.1. Plant Materials Leaves, flowers and rhizomes of A. palaestinum were collected and identified during the spring (March–June, 2022) from the hills of South Lebanon, the plant was identified by the pharmacognosist Dr. Mohamad Khiami and can be found at the herbarium of the Lebanese University. The leaves and flowers were washed with distilled water then dried for 20 days in the shade at room temperature. The rhizomes were washed many times to eliminate the soil particles, cut to small pieces, dried for 40 days in the shade at room temperature. All dried parts were grounded and the powder were stored at 4 °C until their use. 2.2. Preparation of the Extracts Extracts of each part (10%) were prepared using distilled water, ethanol (99.4%) or choloroform (99.4%). Samples were placed in a shaker at 100 rounds per minute for 72 hours at room temperature, then filtered through Whatman filter papers (No.4).The extraction was repeated three times.Thereafter, the combined filtrate were then dried by rotary evaporation and stored at -20°C for later use. Dried yields of ethanolic and chloroformic extracts were dissolved in 4% DMSO and in distilled water for aqueous extracts at a final concentration of 50 mg/mL. 2.3. LC-MS Analysis Separation and detection of the phytochemical compounds from the water, ethanol and chloroform extracts were performed on an AB Sciex X500R QTOF ESI mass spectrometer at the American Unviersity of Beirut. LC flow was split to 500 nL/min before entering the ion source. Mass spectra were acquired in centroid mode ranging from 150 to 1,000 m/z, resolution R = 30,000. A Luna Omega C18, 150 × 2.1 mm, 1.6 µm column was used, with injection volume of 1 µL. A gradient of A) H2O+ 0.1% FA (formic acid) and B) Acetonitrile+0.1% FA at a flow rate of 0.55 mL/min was used to achieve separation. Gradient conditions started at 5%B, increase to 10% B in 1 min, then to 35% B from minute 1→15, then 855 Edelweiss Applied Science and Technology ISSN: 2576-8484 Vol. 9, No. 2: 853-862, 2025 DOI: 10.55214/25768484.v9i2.4619 © 2025 by the authors; licensee Learning Gate to 50% B from minute 15→22, and finally to 80% B from minute 22→25. After a 1-min hold at 80% B, the system was re-equilibrated for 5 min with the initial conditions. UV data was acquired using a PDA (wavelength 200–800 nm ± 8 nm), MS detection was performed simultaneously. 2.4. Determination of Antibacterial Activity of A. Palaestinum Extracts Against P. Syringae Pv. Tomato (Pst) 2.4.1. Activity By Disc Diffusion Method A preliminary check of the antibacterial acitivites of the prepared extract against P. syringae was performed using the disc diffusion method according to Leite, et al. [21]. P. syringae pv. tomato (Pst) at 1.0 x 105 CFU/mL was incubated in King Agar B for 24 h at 28◦C. Discs (6 mm) of Whatmann filter paper (No. 1) were sterilized in an oven at 160 ◦C for one hour. Discs were impregnated with10 μL of the prepared extract, dried under a laminar flow sterile bench. Positive control discs containing 0.5mg of the reference bacteriocide copper oxychloride, 85%WP (Wettable Powders) (Agro Life Science Corporation, India) as (C1), or 0.5 mg Ciproloxacin (C2) (Macleods’ Pharmceuticals LTD, Mumbai, India) as (C2) were used as positive controls while discs containing 10μL of 4% DMSO (C-) were used as negative control. All Petri dishes were sealed to avoid possible evaporation of the test samples, then incubated at 28◦C for 24 h. After incubation, the diameters of the bacterial growth inhibition zones were measured including the disc diameters. The antibacterial activity was expressed as the mean inhibition zone diameters, IZD (mm), and relative antimicrobial activity (RAA). The tests were performed in triplicates. The relative antimicrobial activity was calculated as follows: RAA = [(inhibition zone diameter mean of active plant)2/ (inhibition zone diameter mean of reference antibiotic)2] × 100 2.4.2. Determination of Minimum Inhibitory And Bactericidal Concentrations The minimal inhibitory concentration (MIC) values were determined by broth dilution assay [22]. MIC and minimum bactericidal concentrations (MBC) of the A. palaestinum extracts were determined in nutrient broth using the standard dilution technique. Negative control (tube containing extract and growth medium without inoculum) and positive control (tube containing growth medium inoculated with bacteria) were used. In test tubes, 0.5mL of each diluted extracts with different concentration (5,10,15,20,25,30,35,40,45 and 50 mg/mL) were added to 10 mL nutrient broth inoculated with P. syringae at 105 CFU/mL. All tubes were incubated at 28°C for 48 hours. MIC values were taken as the lowest concentration of extracts that produced no visible bacterial growth when compared with the control tubes. In order to evaluate MBC, 100 μL of each MIC tubes and next tubes with higher concentration of extracts was placed on nutrient agar and incubated at 28°C for 48 hr. The lowest concentration which no bacterial growth observed on nutrient agar plates was considered as MBC. The tests were conducted in triplicates. 2.4.3. Antimicrobial Activity of A. Palaestinum Extracts Against Tomatoes’ Bacterial Speck Disease The plant extracts were evaluated in vivo against bacterial specks in tomatoes. Three tomato plants (8 weeks old) kept inside a greenhouse with 65% relative humidity at 28oC were used for each treatment. The plants were sprayed with 50 mL of P. syringae suspension at 105 CFU/mL. Three days after inoculation, plants were treated with either sterile distilled water (SDW) as negative control, copper oxichloride at 2.5mg/mL and ciproloxacin at 0.5mg/mL dissolved in water as positive controls (C1and C2, respectively) or A. palaestinum extracts (45 mg/mL). After 5 days, 3 leaflets were randomly picked from every tomato plants and the number of specks in each was counted. The means and standard deviations were then caculated. 856 Edelweiss Applied Science and Technology ISSN: 2576-8484 Vol. 9, No. 2: 853-862, 2025 DOI: 10.55214/25768484.v9i2.4619 © 2025 by the authors; licensee Learning Gate 2.5. Statistical Analysis Statistical analyses were performed by Microsoft Excel 2019 to calculate the means and standard deviation (SD). 3. Results 3.1. Identification of Chemical Components of A. Palaestinum Extracts by LC-MS HPLC-MS analysis was based on the peak area percentages, retention time, molecular formula and molecular weight. Thirteen compounds were detected and present in the analysis of aqueous, ethanolic and chloroform fractions of different parts of A. palaestinum (Table 1). It revealed the presence of one chemical compounds in aqueous leaves and flowers’ fractions and 3 in aqueous rhizomes’ fraction, nine other chemical compounds in ethanolic leaves’ extract where 5 were identified in the ethanolic flower extracts and 4 in the ethanolic rhizomes’ extract. In the chloroform extracts, 5 compounds were identified in the leaves and flowers each, while 4 were in the rhizomes. N-Octadecylbenzylamine was common in all aqueous fractions and the two chemical compounds 10,12-pentacosadiynamine and Eicosenamide were found in aqueous rhizomes’ fraction. Palmitamide, Linolenyl alcohol, Octadecanamide and Geranylcitronellol were common in all ethanolic and chloroformic fractions. Pyritiduim, Cyclopiamine A and Ethylpheophorbide A were in the ethanolic leaves’ fraction, Piptamine was common in the ethanolic leaves and flowers and chloroform leaves fractions. Two additional compounds were detected in the ethanolic leaves and chloroformic flowers fractions but were not identified. 857 Edelweiss Applied Science and Technology ISSN: 2576-8484 Vol. 9, No. 2: 853-862, 2025 DOI: 10.55214/25768484.v9i2.4619 © 2025 by the authors; licensee Learning Gate Table 1. LC–MS analysis of the aqueous, ethanolic, and chloroform fractions of A. palaestinum. Fractions m/z (Da) Retention time (min) Molecular formula Identification Aqueous leaves’ extract 360.3607 17.325 C25H45N* N-Octadecylbenzylamine Aqueous flowers’ extract 360.3607 17.459 C25H45N* N-Octadecylbenzylamine Aqueous rhizomes’ extract 326.3770 13.546 C22H47N 10,12-pentacosadiynamine 360.3612 17.464 C25H45N* N-Octadecylbenzylamine 310.3101 15.437 C20H39NO Eicosenamide Ethanolic leaves’ extract Ethanolic leaves’ extract 438.2379 6.287 C26H27N7 Pyritidium 468.2482 6.470 C26H33N3O5 Cyclopiamine A 256.2628 13.663 C16H33NO** Palmitamide 282.2784 13.896 C18H32O** Linolenyl alcohol 593.2741 14.303 C35H63N4O5 Not found in database 284.2941 15.116 C18H37NO** Octadecanamide 310.3095 15.208 C20H36O** Geranylcitronellol 332.3303 15.300 C23H41N*** Piptamine 621.3056 15.861 C37H40N4O5 Ethyl pheophorbide A Ethanolic flowers’ extract 256.2625 13.673 C16H33NO** Palmitamide 282.2778 13.892 C18H32O** Linolenyl alcohol 332.3311 14.834 C23H41N*** Piptamine 284.2939 15.126 C18H37NO** Octadecanamide 310.3095 15.235 C20H36O** Geranylcitronellol Ethanolic rhizomes’’ extract 256.2624 13.651 C16H33NO** Palmitamide 282.2786 13.899 C18H32O** Linolenyl alcohol 284.2944 15.119 C18H37NO** Octadecanamide 310.3101 15.216 C20H36O** Geranylcitronellol Chloroformic leaves’ extract 256.2623 13.665 C16H33NO** Palmitamide 282.2783 13.889 C18H32O** Linolenyl alcohol 284.2941 15.111 C18H37NO** Octadecanamide 310.3097 15.221 C20H36O** Geranylcitronellol 332.3302 13.345 C23H41N*** Piptamine Chloroformic flowers’ extract 537.1024 8.872 C26H21N2O9P Not found in database 256.2628 13.663 C16H33NO** Hexadecanamide 282.2784 13.895 C18H32O** Linolenyl alcohol 284.2937 15.112 C18H37NO** Octadecanamide 310.3099 15.206 C20H36O** Geranylcitronellol Chloroformic rhizomes’extract 256.2628 13.664 C16H33NO** Hexadecanamide 282.2781 13.899 C18H32O** Linolenyl alcohol 284.2941 15.135 C18H37NO** Octadecanamide 310.3099 15.227 C20H36O** Geranylcitronellol Note: * Mean common peaks in the aqueous leaves, flowers and rhizomes fractions. ** Mean common peaks in all ethanolic and chloroform fractions, while *** mean common peaks in ethanolic leaves, ethanolic flowers, and chloroform leaves extracts fractions. 3.2. Antimicrobial Activity of A. Palaestinum The antimicrobial activities of the leaves (L), flowers (F) and rhizomes (R) of A. palaestinum extracts against the phytobacterium P. syringae. using the disk diffusion method is shown in Figure 1 and Table 2. DMSO, the vehicle used to suspend the dried extract that was used as a negative control had no affect on the bacerial growth. 858 Edelweiss Applied Science and Technology ISSN: 2576-8484 Vol. 9, No. 2: 853-862, 2025 DOI: 10.55214/25768484.v9i2.4619 © 2025 by the authors; licensee Learning Gate Figure 1. Representative Figure of Agar Diffusion Assay of A. palaestinum Extracts against P. syringae. L: Leaves extract, F: Flowers extract, R: Rhizomes extract, C1:copper oxichloride , C2 :Ciproloxacin and C- :DMSO. As seen in Figure 1, the aqueous extracts showed the highest antimicorbial activity and was confirmed by the calculated relative antimicrobial activities RAA1 and RAA2 (Table 2). The aqueous extracts yielded a high RAA2 (≥70) with IZD between 26.6 and 28.3mm. This is followed by the ethanolic extracts and chloroform leaves extracts with moderate RAA2 (≥30) and IZD ranging between 17.3 and 20.6mm against P. syringae. The chloroform flowers and rhizomes extracts showed low RAA2 (< 30) and IZD between 15.6 and 15.3mm, respectively. On the other hand, all A. palaestinum extracts showed high RAA1 activity against P. syringae. Table 2. Inhibition zones diameters (IZD) and relative antimicrobial activities to C1 (RAA1) and to C2 (RAA2) of A. palaestinum Extracts against P. syringae. A. palaestinum extracts’ type IZD±SD (mm) RAA1 RAA2 Aqueous leaves 28.3±0.57 408.6 83.3 Aqueous flowers 27.6±0.57 388.6 79.2 Aqueous rhizomes 26.6±0.57 361 73.6 Ethanolic leaves 20.6±0.57 216 44.1 Ethanolic flowers 19.6±0.57 196 39.9 Ethanolic rhizomes 17.6±0.57 158 32.2 Chloroformic leaves 17.3±0.57 152.6 31.14 Chloroformic flowers 16±1 130.6 26.6 Chloroformic rhizomes 15.3±0.57 119.4 24.3 C1 14±0 100 20.3 C2 31±0 490 100 Note: SD: Standard deviation 3.2. MIC and MBC of the Active Plant Extracts The minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of A. palaestinum extracts against P. syringae were determined (Table 3). The phytopathogen was sensitivite to the extracts with MIC and MBC ranging between 10-35 mg/mL and 10-40 mg/mL respectively. These results are in concordance with the inhibitory activities obtained. The aqueous leaves’ extract exhibited by far the strongest bactericidal effects on P. syringae with an MIC and MBC of 10 mg/mL. As the aqueous flowers and rhizomes extracts, MIC of 10mg/mL and a MBC of 15mg/mL were obtained for both. The MIC and MBC of the ethanolic leaves extract were higher (25mg/mL) while the MIC and MBC of the ethanolic flowers and rhizomes extracts were the same, 25 mg/mL and 30 mg/mL 859 Edelweiss Applied Science and Technology ISSN: 2576-8484 Vol. 9, No. 2: 853-862, 2025 DOI: 10.55214/25768484.v9i2.4619 © 2025 by the authors; licensee Learning Gate respecetively. The chloroformic extracts showed the higher values of MIC ranging between 30-35 mg/mL and MBC ranging between 30-40mg/mL. Table 3. MIC and MBC of Arum palaestinum extracts against Pseudomonas syringae. A. palaestinum extracts MIC±SD (mg/mL) MBC±SD (mg/mL) Aqueous leaves 10±0 10±0 Aqueous flowers 10±0 15±5 Aqueous rhizomes 10±0 15±0 Ethanolic leaves 25±5 25±5 Ethanolic flowers 25±0 30±0 Ethanolic rhizomes 25±5 30±0 Chloroformic leaves 30±5 30±0 Chloroformic flowers 35±0 40±0 Chloroformic rhizomes 30±5 35±0 Note: SD: Standard deviation. 3.4. Activity of A. Palaestinum Extracts Against Tomatoe’s Bacterial Speck Disease in Vivo The activity of the different extracts compared to the bacteriocide copper oxychloride (C1) and the antibiotic ciprofloxacin (C2) against tomatoes’ bacterial speck is shown in Table 4. In the field, the effect of A. palaestinum extracts on speck disease was noticeable. The aqueous, ethanolic and chloroform leaves’ extracts were the most powerful as an antibacterial where an obtained average of ˂1 speck/leaflet, while treatment with the chloroform flowers and rhizomes extracts showed means of 9.6 and 10.2 specks/leaflet, respectively. The negative control recorded the highest number of specks of 30.8 specks per leaflet. The positive controls C1 and C2 recorded 4.7 and 0 respectively. Table 4. The effects of A. palaestinum extracts on the bacterial speck disease in the field. C-: negative control (distilled water), C1: positive control 1 (copper oxychloride), C2: positive control 2(Ciprofloxacin). A. palaestinum extracts Mean of number speck/leaflet ±SD Aqueous leaves 0.33±0.50 Aqueous flowers 0.33±0.50 Aqueous rhizomes 0.55±0.52 Ethanolic leaves 0.55±0.52 Ethanolic flowers 0.66±0.50 Ethanolic rhizomes 0.66±0.50 Chloroform leaves 0.88±0.33 Chloroform flowers 9.66±1.30 Chloroform rhizomes 10.22±2.12 C1 4.77±0.83 C2 0.0±0.0 C- 30.88±2.34 Note: SD: Standard deviation. 4. Discussion Arum is a genus of about 26 species of flowering plants in the family Araceae, native to different parts of the globe with the highest species diversity in the Mediterranean region. Around 180 phytochemicals were identified in the leaves of A. palaestinum, mainly flavonoids (quercetin dihexoside, vitexin-O-glucoside,…), phenolic acids (caffeoyl-hexose, pyrocatechol,…) , terpenoids (euphopubescenol, masilinic acid,…), alkaloids, iridoids and amino acids along with unknown compounds [4, 23]. The LC- MS analysis used in this study revealed for the first time the presence of 13 new compounds (N- Octadecylbenzylamine; 10,12-pentacosadiynamine; Eicosenamide; Pyritiduim; Cyclopiamine A; 860 Edelweiss Applied Science and Technology ISSN: 2576-8484 Vol. 9, No. 2: 853-862, 2025 DOI: 10.55214/25768484.v9i2.4619 © 2025 by the authors; licensee Learning Gate Palmitamide; Linolenyl alcohol; Octadecanamide; Geranylcitronellol; Piptamine; Ethyl pheophorbide A and two unknown compounds) in the tested samples of A. palaestinum. Moreover, the LC-MS analysis revealed the presence of chemical components with different biological functions. Three important chemical compounds common in ethanolic and chloroformic extracts of leaves, flowers and rhizomes of A. palaestinum have been discovered, Linolenyl alcohol is described as an antibacterial agent [24] Palmitamide for tissue regeneration preparation [25] and Geranylcitronellol which is an antileishmanial agent [26]. Three other important chemical components of ethanolic leaves’ extract have been detected, Pyritidium acts as antiprotozoal agent and has antiviral properties [27] Cyclopiamine A is a oxindole alkaloid with antitumor and antibacterial properties [28], and Ethyl pheophorbide A has significant anti-proliferative effects in several human cancer cell lines [29] and broad virucidal activities [30]. Furthermore, Piptamine present in the ethanolic leaves’ and flowers’ extracts and chloroformic leaves’ extract also acts as an antibiotic [31] . The antibacterial activity assessment of A. palaestinum in vitro demonstrated that the aqueous and ethanolic extracts of all parts and chloroformic leaves extract had a significant antibacterial activity against P. syringae. Chloroformic flowers and rhizomes extracts further revealed an inhibitory effect on P. syringae. In addition, all extracts showed an inhibitory effect on growth, spread and infection of speck disease on the tomato plants in the field experiment. This results is compatible and related with the LC- MS analysis results, which showed the presence of antibacterial compounds in ethanolic and chloroformic extracts of leaves, flowers and rhizomes of A.palaestinum . Our result is compatible with previous studies reported about the antibacterial effects of Arum subspecies , where it was demonstrated that the crude A. palaestinum flowers aqueous extracts have antibacterial activity against the Gram- positive bacteria Staphylococcus aureus, and Enterococcus faecium; and the Gram-negative bacteria Klebsiella pneumoniae, Proteus vulgaris , Escherichia coli,and Pseudomonas aeruginosa Dwikat, et al. [32]. Jaber, et al. [33] demonstrate that the water and methanol extracts of Arum hygrophilum leaves also had significant antibacterial activity against P. aeruginosa. Also, leaves and berries of Arum maculatum were observed antibacterial activities against P. aeruginosa [34]. The significant antibacterial effect of A. palaestinum against P. syringae reveals a promising hope in their control in organic agriculture like others plant extracts demonstrated by Simonetti, et al. [35] to prevent the bacterial canker caused by p.syringae and use in this disease management. 5. Conclusion This study is considered as a preliminary investigation for future development of naturally occurring bacteriocide to be used in organic agriculture. A. palaestinum extracts revealed promising results to eradicate P. syringae. The plant is rich in phytochemicals worth to be studied in controlling agriculture pests and other agricultural investigations. Transparency: The authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. This study followed all ethical practices during writing. Authors’ Contributions: Jamilah Borjac conceived and designed research. Samaher Ghaith conducted experiments and analyzed the data and wrote the manuscript. Dany Abi Shahine conducted HPLC analyses. Jamilah Borjac, Michel Afram and Antoine Abou Fayad reviewed the manuscript. All authors read and approved the final manuscript. 861 Edelweiss Applied Science and Technology ISSN: 2576-8484 Vol. 9, No. 2: 853-862, 2025 DOI: 10.55214/25768484.v9i2.4619 © 2025 by the authors; licensee Learning Gate Copyright: © 2025 by the authors. This open-access article is distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). References [1] C. L. Henriquez, T. Arias, J. C. Pires, T. B. Croat, and B. A. Schaal, "Phylogenomics of the plant family Araceae," Molecular Phylogenetics and Evolution, vol. 75, pp. 91-102, 2014. https://doi.org/10.1016/j.ympev.2014.02.017 [2] S. Mayo, J. Bogner, E. Catherine, and B. P. Tomlinson, "The genera of Araceae," Kew Bulletin, vol. 53, no. 2, pp. 505- 507, 1998. https://doi.org/10.2307/4114530 [3] H. Migdadi, S. Al-Lozi, I. Makhadmeh, M. Duwayri, and R. Shibli, "Assessment of phenotypic variation of arum species in Jordan," Jordan Journal of Agricultural Sciences, vol. 4, no. 4, pp. 367-379, 2008. https://archives.ju.edu.jo/index.php/jjas/article/view/994 [4] I. M. Abu-Reidah, M. S. Ali-Shtayeh, R. M. Jamous, D. Arráez-Román, and A. Segura-Carretero, "Comprehensive metabolite profiling of Arum palaestinum (Araceae) leaves by using liquid chromatography–tandem mass spectrometry," Food Research International, vol. 70, pp. 74-86, 2015. https://doi.org/10.1016/j.foodres.2015.01.023 [5] S. K. El-Desouky, K. H. Kim, S. Y. Ryu, A. F. Eweas, A. M. Gamal-Eldeen, and Y.-K. Kim, "A new pyrrole alkaloid isolated from Arum palaestinum Boiss. and its biological activities," Archives of Pharmacal Research, vol. 30, pp. 927- 931, 2007. https://doi.org/10.1007/BF02993958 [6] N. Jaradat and M. Abualhasan, "Comparison of phytoconstituents, total phenol contents and free radical scavenging capacities between four Arum species from Jerusalem and Bethlehem," Pharmaceutical Sciences, vol. 22, no. 2, pp. 120- 125, 2016. [7] A. Azab, "Arum: A plant genus with great medicinal potential," European Chemical Bulletin, vol. 6, no. 2, pp. 59-68, 2017. https://doi.org/10.17628/ecb.2017.6.59-68 [8] A. Alsaadi, A. Marwan, A. Myadmah, and M. Kheir, "GC-MS analysis and antioxidant activity of Arum Palaestinum," Dissertation, An-Najah National University. https://repository.najah.edu/server/api/core/bitstreams/f00d003b- 2c6c-4766-abcc-6d68cfcfba0c/content, 2021. [9] A. I. Husein, M. S. Ali-Shtayeh, W. J. Jondi, N. A.-A. Zatar, I. M. Abu-Reidah, and R. M. Jamous, "In vitro antioxidant and antitumor activities of six selected plants used in the Traditional Arabic Palestinian herbal medicine," Pharmaceutical Biology, vol. 52, no. 10, pp. 1249-1255, 2014. https://doi.org/10.3109/13880209.2014.886274 [10] M. M. Farid, S. R. Hussein, and M. M. Saker, "Conservation, genetic characterization, phytochemical and biological investigation of black calla lily: A wild endangered medicinal plant," Asian Pacific Journal of Tropical Disease, vol. 6, no. 10, pp. 832-836, 2016. https://doi.org/10.1016/S2222-1808(16)61141-6 [11] B. Elkhalfi, A. Essari, A. Serrano Delgado, and A. Soukri, "Antibacterial activity of plant methanolic extracts on a field isolate of Pseudomonas syringae pv tomato from the Casablanca region (Morocco)," Advances in Bioscience and Biotechnology, vol. 4, pp. 1-9, 2013. https://doi.org/10.4236/abb.2013.47A2001 [12] S. Nanasombat and P. Lohasupthawee, "Antibacterial activity of crude ethanolic extracts and essential oils of spices against Salmonellae and other enterobacteria," KMITL Science and Technology Journal, vol. 5, no. 3, pp. 527-538, 2005. [13] A. E. Adioumani et al., "Isolation and identification of phytopathogenic bacteria in vegetable crops in West Africa (Côte d'Ivoire)," African Journal of Microbiology Research, vol. 16, no. April, pp. 167–177, 2022. [14] S. Santamaría‐Hernando et al., "Pseudomonas syringae pv. tomato infection of tomato plants is mediated by GABA and l‐Pro chemoperception," Molecular Plant pathology, vol. 23, no. 10, pp. 1433-1445, 2022. [15] X.-F. Xin and S. Y. He, "Pseudomonas syringae pv. tomato DC3000: A model pathogen for probing disease susceptibility and hormone signaling in plants," Annual Review of Phytopathology, vol. 51, no. 1, pp. 473-498, 2013. https://doi.org/10.1146/annurev-phyto-082712-102321 [16] G. M. Preston, "Pseudomonas syringae pv. tomato: The right pathogen, of the right plant, at the right time," Molecular Plant Pathology, vol. 1, no. 5, pp. 263-275, 2000. https://doi.org/10.1046/j.1364-3703.2000.00036.x [17] S. Santamaría-Hernando et al., "The Pseudomonas syringae pv. tomato DC3000 PSPTO_0820 multidrug transporter is involved in resistance to plant antimicrobials and bacterial survival during tomato plant infection," PLoS one, vol. 14, no. 6, p. e0218815, 2019. https://doi.org/10.1371/journal.pone.0218815 [18] C. Cole, T. Burgoyne, A. Lee, L. Stehno-Bittel, and G. Zaid, "Arum Palaestinum with isovanillin, linolenic acid and β- sitosterol inhibits prostate cancer spheroids and reduces the growth rate of prostate tumors in mice," BMC Complementary and Alternative Medicine, vol. 15, no. 1, pp. 1-8, 2015. https://doi.org/10.1186/s12906-015-0774-5 [19] R. Farahmandfar, R. Esmaeilzadeh Kenari, M. Asnaashari, D. Shahrampour, and T. Bakhshandeh, "Bioactive compounds, antioxidant and antimicrobial activities of Arum maculatum leaves extracts as affected by various solvents and extraction methods," Food Science & Nutrition, vol. 7, no. 2, pp. 465-475, 2019. https://doi.org/10.1002/fsn3.815 [20] E. Qnais, Y. Bseiso, M. Wedyan, and H. Alkhateeb, "Evaluation of analgesic activity of the methanol extract from the leaves of Arum palaestinum in mice and rats," Biomedical & Pharmacology Journal, vol. 10, no. 3, p. 1159, 2017. https://dx.doi.org/10.13005/bpj/1216 https://creativecommons.org/licenses/by/4.0/ https://doi.org/10.1016/j.ympev.2014.02.017 https://doi.org/10.2307/4114530 https://archives.ju.edu.jo/index.php/jjas/article/view/994 https://doi.org/10.1016/j.foodres.2015.01.023 https://doi.org/10.1007/BF02993958 https://doi.org/10.17628/ecb.2017.6.59-68 https://repository.najah.edu/server/api/core/bitstreams/f00d003b-2c6c-4766-abcc-6d68cfcfba0c/content https://repository.najah.edu/server/api/core/bitstreams/f00d003b-2c6c-4766-abcc-6d68cfcfba0c/content https://doi.org/10.3109/13880209.2014.886274 https://doi.org/10.1016/S2222-1808(16)61141-6 https://doi.org/10.4236/abb.2013.47A2001 https://doi.org/10.1146/annurev-phyto-082712-102321 https://doi.org/10.1046/j.1364-3703.2000.00036.x https://doi.org/10.1371/journal.pone.0218815 https://doi.org/10.1186/s12906-015-0774-5 https://doi.org/10.1002/fsn3.815 https://dx.doi.org/10.13005/bpj/1216 862 Edelweiss Applied Science and Technology ISSN: 2576-8484 Vol. 9, No. 2: 853-862, 2025 DOI: 10.55214/25768484.v9i2.4619 © 2025 by the authors; licensee Learning Gate [21] S. P. Leite et al., "Antimicrobial activity of Indigofera suffruticosa," Evidence‐Based Complementary and Alternative Medicine, vol. 3, no. 2, pp. 261-265, 2006. https://doi.org/10.1093/ecam/nel010 [22] P. R. Murray, E. J. Baron, M. A. Pfaller, F. C. Tenover, and R. H. Yolke, Manual of clinical microbiology, 6th ed. Washington, DC: American Society for Microbiology Press, 1995. [23] F. U. Afifi, V. Kasabri, S. C. Litescu, and I. M. Abaza, "In vitro and in vivo comparison of the biological activities of two traditionally and widely used Arum species from Jordan: Arum dioscoridis Sibth & Sm. and Arum palaestinum Boiss," Natural Product Research, vol. 30, no. 16, pp. 1777-1786, 2016. https://doi.org/10.1080/14786419.2015.1072713 [24] PubChem, "Linolenyl alcohol - information sources," Retrieved: https://pubchem.ncbi.nlm.nih.gov/compound/Linolenyl-alcohol#section=Information-Sources. [Accessed February 20, 2024], 2024. [25] ChemicalBook, "Chemical product property: CB7262544," Retrieved: https://www.chemicalbook.com/ChemicalProductProperty_EN_CB7262544.htm. [Accessed February 20, 2024], 2024. [26] Chemical Entities of Biological Interest (ChEBI), "CHEBI:24229. European Bioinformatics Institute," Retrieved: https://www.ebi.ac.uk/chebi/searchId.do?chebiId=CHEBI:24229. [Accessed February 20, 2024], 2024. [27] PubChem, "Pyritidium," Retrieved: https://pubchem.ncbi.nlm.nih.gov/compound/Pyritidium. [Accessed 20 February 2024], 2024. [28] E. V. Mercado-Marin et al., "Total synthesis and isolation of citrinalin and cyclopiamine congeners," Nature, vol. 509, no. 7500, pp. 318-324, 2014. https://doi.org/10.1038/nature13273 [29] A. Saide, G. Riccio, A. Ianora, and C. Lauritano, "The Diatom Cylindrotheca closterium and the Chlorophyll Breakdown Product Pheophorbide a for Photodynamic Therapy Applications," Applied Sciences, vol. 13, no. 4, p. 2590, 2023. https://doi.org/10.3390/app13042590 [30] S. Park, J.-Y. Kim, H. C. Kwon, D. S. Jang, and Y.-J. Song, "Antiviral activities of ethyl pheophorbides a and b isolated from Aster pseudoglehnii against influenza viruses," Molecules, vol. 28, no. 1, pp. 1-14, 2022. https://doi.org/10.3390/molecules28010041 [31] PubChem, "Piptamine (antibiotic)," Retrieved: https://pubchem.ncbi.nlm.nih.gov/compound/Piptamine_antibiotic. [Accessed 20 February 2024], 2024. [32] M. Dwikat et al., "Arum palaestinum delays hepatocellular carcinoma proliferation through the PI3K-AKT-mTOR signaling pathway and exhibits anticoagulant effects with antimicrobial properties," Frontiers in Pharmacology, vol. 14, p. 1180262, 2023. https://doi.org/10.3389/fphar.2023.1180262 [33] H. M. Jaber et al., "Antibacterial activity and chemical composition of arum hygrophilum boiss crude extracts," Jordan Journal of Biological Sciences, vol. 13, no. 2, pp. 159–164, 2020. https://jjbs.hu.edu.jo/files/vol13/n2/Paper%20Number%206.pdf [34] N. Erbil, M. Arslan, and Z. T. Murathan, "Antioxidant, antimicrobial, and antimutagenic effects and biochemical contents of Arum maculatum L. That is a medical plant from turkish flora," Fresenius Environmental Bulletin, vol. 12, no. 12/A, pp. 8709-8714, 2018. [35] G. Simonetti et al., "In vitro antimicrobial activity of plant extracts against Pseudomonas syringae pv. actinidiae causal agent of bacterial canker in kiwifruit," Plant Biosystems-An International Journal Dealing with all Aspects of Plant Biology, vol. 154, no. 1, pp. 100-106, 2020. https://doi.org/10.1080/11263504.2019.1699194 https://doi.org/10.1093/ecam/nel010 https://doi.org/10.1080/14786419.2015.1072713 https://pubchem.ncbi.nlm.nih.gov/compound/Linolenyl-alcohol#section=Information-Sources https://www.chemicalbook.com/ChemicalProductProperty_EN_CB7262544.htm https://www.ebi.ac.uk/chebi/searchId.do?chebiId=CHEBI:24229 https://pubchem.ncbi.nlm.nih.gov/compound/Pyritidium https://doi.org/10.1038/nature13273 https://doi.org/10.3390/app13042590 https://doi.org/10.3390/molecules28010041 https://pubchem.ncbi.nlm.nih.gov/compound/Piptamine_antibiotic https://doi.org/10.3389/fphar.2023.1180262 https://jjbs.hu.edu.jo/files/vol13/n2/Paper%20Number%206.pdf https://doi.org/10.1080/11263504.2019.1699194